Insect-borne infectious diseases such as malaria affect at least 1 billion people worldwide, and over 1.5 million of these infected individuals die each year. Malarial parasites and many other vector-borne pathogens have developed resistance to current drugs, and vaccines are not available or feasible for many of these pathogens. Insect vector control programs have historically been an effective strategy for reducing disease transmission. However, some of these diseases have remained intractable while others are experiencing a resurgence. The long-term goal of this project is to identify new safe, selective, and effective insecticides that can be incorporated into an integrated approach for insect vector management. This project will exploit naturally occurring insect-specific pesticidal compounds isolated from the venom of the Australian funnel-web spider Hadronyche versuta. Individual peptide toxins will be used as probes to identify genetically validated insecticide targets that can be moved into an insecticide discovery program. The specific aims for phase I of this project are; A) to continue the functional analysis of three toxins exhibiting a high degree of insect selectivity; B) perform genetic screens with transgenic Drosophila melanogaster lines carrying peptide toxin-encoding genes to identify genetic loci conferring target-site resistance to transgenically-expressed toxins; C) establish stably transformed insect cell lines expressing target genes for use in phase II chemical screens. This multidisciplinary approach will identify genetically validated pesticide targets, which will form the basis of a novel high-throughput screen for safe and selective insecticides in the phase II and III stages of this study.